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International audience; Background: Mucus is known to play a pathogenic role in muco-obstructive lung diseases, but little is known about the determinants of mucus rheology. The purpose of this study is to determine which sputum components influence sputum rheology in patients with muco-obstructive lung diseases.Methods: We performed a cross sectional prospective cohort study. Spontaneous sputum was collected from consecutive patients with muco-obstructive lung diseases. Sputum rheology was assessed using the Rheomuco® rheometer (Rheonova, Grenoble); the elastic modulus G′, viscous modulus G″, and the critical stress threshold σc were recorded. Key quantitative and qualitative biological sputum components were determined by cytology, nucleic acid amplification tests and mass spectrometry.Results: 48 patients were included from January to August 2019. Among them, 10 had asthma, 14 COPD and 24 non-CF bronchiectasis (NCFB). The critical stress threshold σc predicted a sputum eosinophilia superior to 1.25% with 89.19% accuracy (AUC = 0.8762). G′ and G″ are positively correlated with MUC5AC protein concentration ((rho = 0.361; P = .013) and (rho = 0.335; P = .021), respectively). σc was positively correlated with sputum eosinophilia (rho = 0.394; P = .012), MUC5B (rho = 0.552; P < .001) and total protein (rho = 0.490; P < .001) concentrations. G′ and G″ were significantly higher in asthma patients (G’ = 14.49[7.18–25.26]Pa, G’‘ = 3.0[2.16–5.38]Pa) compared to COPD (G’ = 5.01[2.94–6.48]Pa, P = .010; G’‘ = 1.45[1.16–1.94]Pa, P = .006) and to NCFB (G’ = 4.99[1.49–10.49]Pa, P = .003; G’’ = 1.46[0.71–2.47]Pa, P = .002).Conclusion: In muco-obstructive lung diseases, rheology predicts sputum eosinophilia and is correlated with mucin concentrations, regardless of the underlying disease.Clinical trial registration: (registrar, website, and registration number), where applicable NCT04081740.

International audience; As opposed to surface marker staining, certain cell types can only be recognized by intracellular markers. Intracellular staining for use in cell sorting remains challenging. Fixation and permeabilization steps for intracellular staining and the presence of RNases notably affect preservation of high-quality mRNA. We report the work required for the optimization of a successful protocol for microarray analysis of intracellular target-sorted, formalin-fixed human bronchial club cells. Cells obtained from differentiated air-liquid interface cultures were stained with the most characteristic intracellular markers for club cell (SCGB1A1+) sorting. A benchmarked intracellular staining protocol was carried out before flow cytometry. The primary outcome was the extraction of RNA sufficient quality for microarray analysis as assessed by Bioanalyzer System. Fixation with 4% paraformaldehyde coupled with 0.1% Triton/0.1% saponin permeabilization obtained optimal results for SCGB1A1 staining. Addition of RNase inhibitors throughout the protocol and within the appropriate RNA extraction kit (Formalin-Fixed-Paraffin-Embedded) dramatically improved RNA quality, resulting in samples eligible for microarray analysis. The protocol resulted in successful cell sorting according to specific club cell intracellular marker without using cell surface marker. The protocol also preserved RNA of sufficient quality for subsequent microarray transcriptomic analysis, and we were able to generate transcriptomic signature of club cells.

Strawberry (Fragaria × ananassa) fruits are an excellent source of L-ascorbic acid (AsA), a powerful antioxidant for plants and humans. Identifying the genetic components underlying AsA accumulation is crucial for enhancing strawberry nutritional quality. Here, we unravel the genetic architecture of AsA accumulation using an F1 population derived from parental lines ‘Candonga’ and ‘Senga Sengana’, adapted to distinct Southern and Northern European areas. To account for environmental effects, the F1 and parental lines were grown and phenotyped in five locations across Europe (France, Germany, Italy, Poland and Spain). Fruit AsA content displayed normal distribution typical of quantitative traits and ranged five-fold, with significant differences among genotypes and environments. AsA content in each country and the average in all of them was used in combination with 6,974 markers for quantitative trait locus (QTL) analysis. Environmentally stable QTLs for AsA content were detected in linkage group (LG) 3A, LG 5A, LG 5B, LG 6B and LG 7C. Candidate genes were identified within stable QTL intervals and expression analysis in lines with contrasting AsA content suggested that GDP-L-Galactose Phosphorylase FaGGP(3A), and the chloroplast-located AsA transporter gene FaPHT4;4(7C) might be the underlying genetic factors for QTLs on LG 3A and 7C, respectively. We show that recessive alleles of FaGGP(3A) inherited from both parental lines increase fruit AsA content. Furthermore, expression of FaGGP(3A) was two-fold higher in lines with high AsA. Markers here identified represent a useful resource for efficient selection of new strawberry cultivars with increased AsA content.

L’impact en santé publique des pathologies respiratoires chroniques ne cesse de croître. Dans ce contexte, il paraît indispensable d’améliorer les modèles d’études du poumon afin de reproduire au plus proche l’architecture pulmonaire complexe, garante des fonctions d’oxygénation et d’épuration du gaz carbonique. Les connaissances actuelles en physiopathologie respiratoire résultent en partie des études de modèles de reconstitution d’épithélium bronchique in vitro à partir de cellules primaires, en deux dimensions sur des inserts, ou en trois dimensions, en organoïdes mimant jusqu’à l’arborescence pulmonaire. Le développement de ces modèles in vitro a connu un nouvel essor grâce aux organoïdes pulmonaires issus de cellules souches pluripotentes et la démocratisation des outils d’édition du génome. Ces apports technologiques récents offrent de nouvelles perspectives en matière de thérapeutiques ou de compréhension physiopathologique et pourraient, dans le futur, ouvrir les portes de la médecine régénératrice pulmonaire.

Fruit colour is central to the sensorial and nutritional quality of strawberry fruit and is therefore a major target in breeding programmes of the octoploid cultivated strawberry (Fragaria × ananassa). The red colour of the fruit is caused by the accumulation of anthocyanins, which are water-soluble flavonoids. To facilitate molecular breeding, here we have mapped with high resolution fruit colour quantitative trait loci (QTLs) (COLOUR, scored visually as in selection programmes) and associated flavonoid metabolic QTLs (5 anthocyanins compounds together with 8 flavonols and flavan-3-ols) to specific subgenomes of cultivated strawberry. Two main colour-related QTLs were located on the LG3A linkage group (Fragaria vesca subgenome). Genetic mapping, transcriptome analysis and whole genome sequencing enabled the detection of a homoeo-allelic variant of ANTHOCYANIDIN REDUCTASE (ANR) underlying the major male M3A COLOUR and pelargonidin-3-glucoside (PgGs) QTLs (up to ∼20% of explained variance). Consistent with previously published functional studies, ANR transcript abundance was inversely related with PgGs content in contrasted progeny individuals. Genetic segregation analyses further indicated that a molecular marker designed using an 18 bp deletion found in the 5′UTR of the candidate ANR homoeo-allelic variant is effective in identifying genotypes with intense red fruit colour. Our study provides insights into the genetic and molecular control of colour-related traits in strawberry and further defines a genetic marker for marker-assisted selection of new strawberry varieties with improved colour. The QTLs detected and the underlying candidate genes are different from those described to date, emphasising the importance of screening a wide diversity of genetic resources in strawberry.

The effect of cultivar and environmental variations and their interaction on anthocyanin components of strawberry were assessed for six cultivars grown in five locations from North to South of Europe in two different years. To evaluate the impact of latitude- and altitude-related factors, daily mean (Tmean), maximum (Tmax) and minimum (Tmin) temperature and global radiation accumulated for 3, 5, 10 and 15 days before fruit sampling, was analyzed. In general, fruits grown in the south were more enriched in total anthocyanin and pelargonidin-3-glucoside (pel-3-glc), the most abundant anthocyanin in strawberry. Principal component analysis (PCA) provided a separation of the growing locations within a cultivar due to latitudinal climatic differences, temporary weather changes before fruit collection and cultivation technique. PCA also depicted different patterns for anthocyanin distribution indicating a cultivar specific reaction on the environmental factors. The linear regression analysis showed that pel-3-glc was relatively less affected by these factors, while the minor anthocyanins cyanidin-3-glucoside, cyanidin-3-(6-O-malonyl)-glucoside, pelargonidin-3-rutinoside and pelargonidin-3-(6-O-malonoyl)-glucoside were sensitive to Tmax. The global radiation strongly increased cya-3-mal-glc in ‘Frida’ and pel-3-rut in ‘Frida’ and ‘Florence’. ‘Candonga’ accumulated less pel-3-glc and total anthocyanin with increased global radiation. The anthocyanin profiles of ‘Gariguette’ and ‘Clery’ were unaffected by environmental conditions.

RationaleHighly reproducible in vitro generation of human bronchial epithelium from pluripotent stem cells is an unmet key goal for drug screening to treat lung diseases. The possibility of using induced pluripotent stem cells (hiPSC) to model normal and diseased tissue in vitro from a simple blood sample will reshape drug discovery for chronic lung, monogenic and infectious diseases.MethodsWe devised a simple and reliable method that drives a blood sample reprogrammed into hiPSC subsequently differentiated within 45 days into air-liquid interface bronchial epithelium (iALI), through key developmental stages, definitive-endoderm (DE) and Ventralized-Anterior-Foregut-Endoderm (vAFE) cells.ResultsReprogramming blood cells from one healthy and 3 COPD patients, and from skin-derived fibroblasts obtained in one PCD patient, succeeded in 100% of samples using Sendai viruses. Mean cell purity at DE and vAFE stages was greater than 80%, assessed by expression of CXCR4 and NKX2.1, avoiding the need of cell sorting. When transferred to ALI conditions, vAFE cells reliably differentiated within 4 weeks into bronchial epithelium with large zones covered by beating ciliated, basal, goblets, club cells and neuroendocrine cells as found in vivo. Benchmarking all culture conditions including hiPSCs adaptation to single-cell passaging, cell density and differentiation induction timing allowed for consistently producing iALI bronchial epithelium from the five hiPSC lines.ConclusionsReliable reprogramming and differentiation of blood-derived hiPSCs into mature and functional iALI bronchial epithelium is ready for wider use and this will allow better understanding lung disease pathogenesis and accelerating the development of novel gene therapies and drug discovery.

Fruit colour, which is central to the sensorial and nutritional quality of the fruit, is a major breeding target in cultivated strawberry (Fragaria × ananassa). Red fruit colour is caused by the accumulation of anthocyanins, which are water-soluble flavonoids. Here, using pseudo F1progeny derived from the cv. ‘Capitola’ and the advanced line ‘CF1116’ and taking advantage of the available high density SNP array, we delineated fruit flavonoids mQTLs (13 compounds: anthocyanin, flavonols and flavan-3-ols) and colour-related QTLs (total anthocyanins and fruit colour) to narrow genomic regions corresponding to specific subgenomes of the cultivated strawberry. Findings showed that the overwhelming majority of the anthocyanin mQTLs and other colour-related QTLs are specific toF. vescasubgenome but that other subgenomes also contribute to colour variations. We then focused on two major homoeo-mQTLs for pelargonidin-3-glucoside (PgGs) localized on both male and female maps on linkage group LG3a (F. vescasubgenome). Combined high-resolution mapping of PgGs mQTLs and colour QTLs, transcriptome analysis of selected progeny individuals and whole genome sequencing of the parents led to the identification of several INDELS in thecis-regulatory region of aMYB102-like ODORANTgene and of the deletion of a putative MADS box binding motif in the 5’UTR upstream region of an anthocyanidin reductase (ANR)gene, which likely underlie significant colour variations in strawberry fruit. The implications of these findings are important for the functional analysis and genetic engineering of colour-related genes and for the breeding by Marker-Assisted-Selection of new strawberry varieties with improved colour and health-benefits.

Flavonoids are products from specialized metabolism that contribute to fruit sensorial (color) and nutritional (antioxidant properties) quality. Here, using a pseudo full-sibling F

L’animation pornographique et érotique commerciale semble avoir toujours existé sur les écrans français. Son histoire commence par les films courts d’animation destinés aux maisons closes du début du XXe siècle, en passant par les grands succès commerciaux de longs-métrages dans les années 1970 tels que La Honte de la jungle ou Fritz le chat, jusqu’à la diffusion estivale en 2018 de la série d’animation française Peepoodo and the Super Fuck Friends. Pour autant, cette production subit une marginalisation économique, juridique et morale, limitant sa distribution. À partir d’une rétrospective historique puis d’une étude de cas, cet article se propose d’apporter une première réponse à la question de savoir si une industrie de l’animation pornographique – et non pas seulement un ensemble de contenus cinématographiques et audiovisuels – aurait la possibilité d’exister dans la France des années 2010. Pornographic and erotic commercial animation seems to have always existed on French screens. Its history begins from short animated movies shown in brothels in early 20th century, to big commercial successes of long animated features in the 1970s such as Tarzoon or Fritz the Cat, until the summer distribution of the French animated series Peepoodo and the Super Fuck Friends in 2018. However, this production suffers from an economic, legal and moral marginalization, restraining its distribution. Through historical retrospective and a case study, this article proposes to bring an answer to the question of the possible existence of a pornographic and erotic animation industry in France in the 2010s.

Flavonoids are products from specialized metabolism that contribute to fruit sensorial (color) and nutritional (antioxidant properties) quality. Here, using a pseudo full-sibling F1 progeny previously studied for fruit sensorial quality of cultivated strawberry (Fragaria × ananassa), we explored over two successive years the genetic architecture of flavonoid-related traits using liquid chromatography electrospray ionization tandem mass spectrometry (13 compounds including anthocyanins, flavonols, and flavan-3-ols) and colorimetric assays (anthocyanins, flavonoids, phenolics, and total antioxidant capacity (ferric reducing antioxidant power and Trolox equivalent antioxidant capacity)). Network correlation analysis highlighted the high connectivity of flavonoid compounds within each chemical class and low correlation with colorimetric traits except for anthocyanins. Mapping onto the female and male linkage maps of 152 flavonoid metabolic quantitative trait loci (mQTLs) and of 26 colorimetric QTLs indicated colocalization on few linkage groups of major flavonoid- and taste-related QTLs previously uncovered. These results pave the way for the discovery of genetic variations underlying flavonoid mQTLs and for marker-assisted selection of strawberry varieties with improved sensorial and nutritional quality.

Summary Background Indicators of the appropriateness of oral anticoagulant prescriptions are lacking, despite the major contribution they could make to improve quality of care. Aim To identify and select such indicators according to their utility and operational implementation. Methods A literature review was conducted to identify indicators of the appropriateness of oral anticoagulant prescriptions according to the guidelines of health authorities and European learned societies. A first list of indicators was identified from guidelines related to general or targeted clinical situations. A two-round Delphi consensus process, completed by a synthesis meeting, was then set up to ask European experts to rate the utility and operational implementation of the indicators on a qualitative binary scale. An indicator was selected if ≥ 80% of the experts judged it both useful and implementable (strong consensus). Results We selected 32 references, from which 84 indicators were identified. Nineteen indicators were short-listed for submission to expert judgment. Twenty-two experts participated in the Delphi process. Sixteen indicators obtained strong consensus for selection; three indicators did not achieve consensus. Two-thirds of the selected indicators focused on the appropriateness of oral anticoagulant prescriptions in general or in patients with atrial fibrillation; the other third focused on the appropriateness of prescriptions in patients with a prosthetic heart valve, venous thromboembolism or trauma. Conclusion This work addresses the current lack of indicators of the appropriateness of oral anticoagulant prescriptions. The selected indicators will be implemented from the hospital information system to assess their metrological properties to detect inappropriate prescriptions.

The airway epithelium represents a fragile environmental interface potentially disturbed by cigarette smoke (CS), the major risk factor for developing chronic obstructive pulmonary disease (COPD). CS leads to bronchial epithelial damage on ciliated, goblet, and club cells, which could involve calcium (Ca

Club cell secretory protein (CCSP) knockout mice exhibit increased airway neutrophilia, as found in chronic obstructive pulmonary disease (COPD). We therefore investigated whether treating COPD airway epithelia with recombinant human CCSP (rhCCSP) could dampen exaggerated airway neutrophilia.Control, smoker and COPD air–liquid interface (ALI) cultures exposed to cigarette smoke extract (CSE) were treated with and without rhCCSP. The chemotactic properties of the supernatants were assessed using Dunn chambers. Neutrophil chemotaxis along recombinant human interleukin 8 (rhIL8) gradients (with and without rhCCSP) was also determined. rhCCSP–rhIL8 interactions were tested through co-immunoprecipitation, Biacore surface plasmon resonance (SPR) andin silicomodelling. The relationship between CCSP/IL8 concentration ratios in the supernatant of induced sputum from COPD patientsversusneutrophilic airway infiltration assessed in lung biopsies was assessed.Increased neutrophilic chemotactic activity of CSE-treated ALI cultures followed IL8 concentrations and returned to normal when supplemented with rhCCSP. rhIL8-induced chemotaxis of neutrophils was reduced by rhCCSP. rhCCSP and rhIL8 co-immunoprecipitated. SPR confirmed thisin vitrointeraction (equilibrium dissociation constant=8 µM).In silicomodelling indicated that this interaction was highly likely. CCSP/IL8 ratios in induced sputum correlated well with the level of small airway neutrophilic infiltration (r2=0.746, pCCSP is a biologically relevant counter-balancer of neutrophil chemotactic activity. These different approaches used in this study suggest that, among the possible mechanisms involved, CCSP may directly neutralise IL8.

To prevent extravasations in medical imaging, analyses of organizational and professional circumstances of the occurrence of extravasations have been conducted in the Bordeaux University Hospital (BUH).Two parallel studies were conducted: (i) clinical practice evaluation (CPE) including practice analysis groups and development of indicator of extravasation occurrence; (ii) Case-control study analyzing the factors associated with the occurrence of extravasations. Cases were constituted with consecutive series of patients with extravasation occurred in the BUH; controls were constituted of series of patients managed in medical imaging in the BUH without any extravasation. Statistical analysis included univariate and multivariate regression logistic models.CPE identified the following circumstances: patient' characteristics (female gender, low blood vessels) and professional situations (lack of check of: peripheral veinous access, contrast media temperature, injection process and lack of matching between injection amount and veinous access diameter). In the case-control analysis, patients' gender and catheter placement in medical imaging were significantly associated with the occurrence of extravasations. Guidelines have been drafted allowing the decrease of extravasation incidence.Adherence to guidelines of contrast media preparation and injection process is very important to prevent extravasation and improve patient safety.

There is a growing interest in developing tailored non-pharmacological strategies to face patients’ needs in dementia. Occupational therapy (OT) may contribute to promote self-empowerment of both patients and caregivers. France has implemented nationwide OT over a short-term period of 3/4 months. The main objective of the MatheoAlz study is to measure the impact of maintaining OT over 4 supplementary months on patients’ neuropsychiatric symptoms. The MatheoAlz trial (Maintenance of Occupational Therapy in Alzheimer’s disease) is a multi-center, pragmatic randomized controlled trial testing maintenance of OT over 4 supplementary months compared to routine OT delivered as recommended. This paper describes the study protocol. MatheoAlz plans to enroll 240 dyads, i.e. dementia patients and caregivers, whose main inclusion criteria are: prescription for routine OT, patients with mild or moderate dementia, living at home, receiving support from an informal caregiver. The study will compare a control group of patients benefiting from 12 to 15 initial sessions of OT over 3/4 months and an intervention group of patients benefiting from these initial sessions plus 8 extra home sessions over 4 supplementary months. The main outcome is the patient’s neuropsychiatric symptoms assessed by the Neuropsychiatric Inventory at 8 months. Several clinical outcomes and economic consequences are measured at 4, 8 and 12 months. This is the first trial designed to assess the specific impact of the maintaining OT on the patients’ neuropsychiatric symptoms burden. The results will inform policymakers on strategies to implement in the near future. This trial was registered at ClinicalTrials.gov on February 16, 2018, identifier: NCT03435705 .

International audience

We describe an original method to measure mucus microrheology on human bronchial epithelium culture using optical tweezers. We probed rheology on the whole thickness of mucus above the epithelium and showed that mucus gradually varies in rheological response, from an elastic behavior close to the epithelium to a viscous one far away. Microrheology was also performed on mucus collected on the culture, on ex vivo mucus collected by bronchoscopy, and on another epithelium model. Differences are discussed and are related to mucus heterogeneity, adhesiveness, and collection method.

Introduction : Les circonstances de survenue des extravasations de produit de contraste en imagerie au CHU de Bordeaux ont ete analysees de facon a pouvoir definir les leviers de leur prevention.Methode : Ont ete mises en place : (i) une evaluation des pratiques professionnelles (EPP) s’appuyant sur des groupes d’analyses de pratiques (GAP) et un suivi d’indicateur d’incidence des extravasations ; (ii) une etude cas-temoins analysant les facteurs associes a la survenue des extravasations. Les cas etaient constitues d’une serie consecutive de patients avec signalement d’extravasation, les « temoins », d’une serie consecutive de patients ayant beneficie d’un acte d’imagerie avec injection de produit de contraste, sans survenue d’extravasation. Des modelisations par regressions logistiques univariee et multivariee ont ete realisees.Resultats : Les circonstances identifiees par l’EPP comme favorisant la survenue des extravasations etaient cliniques (sexe feminin et faiblesse du reseau de voies veineuses) et professionnelles (manque de verification de la voie veineuse peripherique [VVP] et de la temperature du produit de contraste, manque d’adequation entre debit d’injection et calibre des VVP, manque de verification du bon deroule de l’injection). L’etude cas-temoins a montre que les variables significativement associees a la probabilite de survenue de l’extravasation etaient le sexe feminin et le lieu de pose de la VVP en imagerie. Des recommandations ont ete elaborees dont la diffusion a permis de diminuer l’incidence des extravasations.Conclusion : Le respect scrupuleux des recommandations de preparation du produit de contraste et de conduite de l’injection est essentiel pour la prevention des extravasations et la securite du patient.

Introduction La production in vitro, de maniere reproductible et en larges quantite, d’un epithelium bronchique a partir de cellules souches pluripotentes est un objectif cle pour le criblage de medicaments a haut debit pour identifier de nouvelles molecules et la production de cellules pour une therapie cellulaire en pneumologie (Pharmacol Ther 2018 :183, 58-77). Methodes Nous avons concu une methode simple et reproductible qui conduit un simple echantillon de sang en hiPSC par reprogrammation, puis differencie en 45 jours en epithelium bronchique en interface air-liquide (iALI), en suivant successivement les etapes cles du developpement que sont la formation de l’endoderme definitif (DE) et de l’endoderme anterieur ventralise (vAFE). Resultats La reprogrammation des cellules sanguines d’un patient sain et de 3 patients atteints de BPCO, ainsi que de fibroblastes derives de la peau obtenus chez un patient PCD, a ete obtenue en utilisant des virus Sendai. La purete moyenne des cellules aux stades DE et vAFE evaluee par l’expression de CXCR4 et NKX2.1, respectivement, etait superieure a 85 %, evitant ainsi la necessite d’un tri cellulaire. Lorsqu’elles sont transferees dans des conditions de culture d’interface air-liquide (ALI), les cellules vAFE se differencient en 4 semaines en epithelium bronchique avec de grandes zones couvertes par les principales sous-populations cellulaires de l’epithelium bronchique : cellules basales (KRT5 + ), ciliees (TubIV + ), a mucus (Muc5AC + ), Club (CCSP + ), ainsi que des cellules neuroendocrines (CHGA + )(epithelium bronchique iALI). La microscopie electronique confirme la presence de cils motiles et leur frequence de battement est similaire a celui observe dans des HBEC. Certains iALI ont ete maintenus pendant plus de 400 jours. Conclusions La reprogrammation et la differenciation fiables des hiPSC derives du sang, quel que soit leur contexte clinique, en epithelium bronchique iALI mature et fonctionnel est maintenant accessible a une utilisation plus large, ce qui permettra de mieux comprendre la pathogenese des maladies pulmonaires et d’accelerer le developpement de nouvelles therapies geniques et la decouverte de medicaments.

Background The appropriateness of psychotropic prescriptions in the elderly is a major quality-of-care challenge at hospital. Quality indicators have been developed to prevent inappropriate psychotropic prescriptions. We aimed to select and automatically calculate such indicators, from the Bordeaux University Hospital information system, and to analyze the appropriateness of psychotropic prescription practices, in an observational study. Methods Experts selected indicators of the appropriateness of psychotropic prescriptions in hospitalized elderly patients, according to guidelines from the French High Authority for Health. The indicators were reformulated to focus on psychotropic administrations. The automated calculation of indicators was analyzed by comparing their measure to data collected from a clinical audit. In elderly patients hospitalized between 2014 and 2015, we then analyzed the evolution of the appropriateness of psychotropic prescription practices during hospital stay, using methods of visualization, and described practices by considering patients’ characteristics. Results Two indicators were automated to detect overuse and misuse of psychotropic drugs. Indicators identified frequent inappropriate drug administrations, but practices tended to become more appropriate after quality-of-care improvement actions. In the majority of patients (85%), there was no inappropriate administration of psychotropic drugs during hospital stay; for the remaining 15% with at least one inappropriate administration, physicians tended to limit overuse or misuse during hospital stay. Inappropriate administrations were more frequent in patients suffering from psychiatric disorders, dependence and associated complications or morbidities. Conclusions The automated indicators are structuring tools for the development of a drug prescription monitoring system. Inappropriate psychotropic administrations were limited by physicians during hospital stay; some inappropriate prescriptions might be explained by clinical characteristics of patients. Electronic supplementary material The online version of this article (10.1186/s12913-019-4064-7) contains supplementary material, which is available to authorized users.

International audience; Poor asthma control and recurrent exacerbations have been shown to be a phenotypic counterpart of asthma with predominantly small-airway involvement.1 Biomarkers are not always accurate in asthmatic patients, especially in serum, because compartmentalization can occur between the blood and airways. Blood eosinophil counts do not represent an overall view of airway inflammation, and exhaled nitric oxide measurements at different flow rates (fraction of exhaled nitric oxide [Feno] and alveolar nitric oxide [Calvno]) have been developed and validated to reflect more accurately proximal and distal airway inflammation.2Club cell secretory protein (CCSP) serum concentration has been shown to be associated with chronic obstructive pulmonary disease, bronchiolitis obliterans syndrome, and sarcoidosis, which are all predominantly diseases involving the small airways. Ranges of CCSP concentrations in healthy subjects, reproducibility, and relationships between serum and airway levels are known and can be used as potential surrogate markers. Our aim was to assess small-airway disease in asthmatic patients and to find a related biomarker. We used a dynamic assessment of gas trapping using computed tomographic (CT) imaging of the chest during methacholine challenge as a marker of small-airway disease.

International audience; The development of suitable Cystic Fibrosis (CF) models for preclinical bench tests of therapeutic candidates is challenging. Indeed, the validation of molecules to rescue the p.Phe508del-CFTR channel (encoded by the Cystic Fibrosis Transmembrane conductance Regulator gene carrying the p.Phe508del mutation) requires taking into account their overall effects on the epithelium. Suberoylanilide Hydroxamic Acid (SAHA), a histone deacetylase inhibitor (HDACi), was previously shown to be a CFTR corrector via proteostasis modulation in CFTR-deficient immortalized cells. Here, we tested SAHA effects on goblet cell metaplasia using an ex vivo model based on the air-liquid interface (ALI) culture of differentiated airway epithelial cells obtained by nasal scraping from CF patients and healthy controls. Ex vivo epithelium grew successfully in ALI cultures with significant rise in the expression of CFTR and of markers of airway epithelial differentiation compared to monolayer cell culture. SAHA decreased CFTR transcript and protein levels in CF and non-CF epithelia. Whereas SAHA induced lysine hyperacetylation, it did not change histone modifications at the CFTR promoter. SAHA reduced MUC5AC and MUC5B expression and inhibited goblet epithelial cell differentiation. Similar effects were obtained in CF and non-CF epithelia. All the effects were fully reversible within five days from SAHA withdrawal. We conclude that, ex vivo, SAHA modulate the structure of airway epithelia without specific effect on CFTR gene and protein suggesting that HDACi cannot be useful for CF treatment.

Background Bronchial epithelium plays a key role in orchestrating innate and adaptive immunity. The fate of ex vivo airway epithelial cultures growing at the air liquid interface (ALI) derived from human endobronchial biopsies or brushings is not easy to predict. Calibrating and differentiating these cells is a long and expensive process requiring rigorous expertise. Pinpointing factors associated with ALI culture success would help researchers gain further insight into epithelial progenitor behavior. Methods A successful ALI culture was defined as one in which a pseudostratified epithelium has formed after 28 days in the presence of all differentiated epithelial cell types. A 4-year prospective bi-center study was conducted with adult subjects enrolled in different approved research protocols. Results 463 consecutive endobronchial biopsies were obtained from normal healthy volunteers, healthy smokers, asthmatic patients and smokers with COPD. All demographic variables, the different fiber optic centers and culture operators, numbers of endo-bronchial biopsies and the presence of ciliated cells were carefully recorded. Univariate and multivariate models were developed. A stepwise procedure was used to select the final logistic regression model. ALI culture success was independently associated with the presence of living ciliated cells within the initial biopsy (OR = 2.18 [1.50–3.16], p

International audience; Impaired airway homeostasis in chronic obstructive pulmonary disease (COPD) could be partly related to club cell secretory protein (CCSP) deficiency. We hypothesize that CCSP G38A polymorphism is involved and aim to examine the influence of the CCSP G38A polymorphism on CCSP transcription levels and its regulatory mechanisms. CCSP genotype and CCSP levels in serum and sputum were assessed in 66 subjects with stable COPD included in a 1-yr observational study. Forty-nine of them had an exacerbation. In an in vitro study, the impact on the CCSP promoter of 38G wild-type or 38A variant was assessed. BEAS-2B cells were transfected by either the 38G or 38A construct, in the presence/absence of cigarette smoke extract (CSE) or lipopolysaccharides (LPS). Cotransfections with modulating transcription factors, p53 and Nkx2.1, identified by in silico analysis by using ConSite and TFSEARCH were conducted. A allele carrier COPD patients had lower serum and sputum CCSP levels, especially among active smokers, and a decreased body mass index, airflow obstruction, dyspnea, and exercise capacity (BODE) score. In vitro, baseline CCSP transcription levels were similar between the wild and variant constructs. CSE decreased more profoundly the CCSP transcription level of 38A transfected cells. The opposite effect was observed with p53 cotransfection. LPS stimulation induced CCSP repression in 38A promoter transfected cells. Cotransfection with Nkx2.1 significantly activated the CCSP promoters irrespective of the polymorphism. Circulating CCSP levels are associated with smoking and the CCSP G38A polymorphism. CSE, LPS, and the Nkx2.1 and p53 transcription factors modulated the CCSP promoter efficiency. The 38A polymorphism exaggerated the CCSP repression in response to p53 and CSE.

National audience

Fruit quality traits are major breeding targets in the Rosaceae. Several of the major Rosaceae species are current or ancient polyploids. To dissect the inheritance of fruit quality traits in polyploid fleshy fruit species, we used a cultivated strawberry segregating population comprising a 213 full-sibling F1 progeny from a cross between the variety ‘Capitola’ and the genotype ‘CF1116’. We previously developed the most comprehensive strawberry linkage map, which displays seven homoeology groups (HG), including each four homoeology linkage groups (Genetics 179:2045–2060, 2008). The map was used to identify quantitative trait loci (QTL) for 19 fruit traits related to fruit development, texture, colour, anthocyanin, sugar and organic acid contents. Analyses were carried out over two or three successive years on field-grown plants. QTL were detected for all the analysed traits. Because strawberry is an octopolyploid species, QTL controlling a given trait and located at orthologous positions on different homoeologous linkage groups within one HG are considered as homoeo-QTL. We found that, for various traits, about one-fourth of QTL were putative homoeo-QTL and were localised on two linkage groups. Several homoeo-QTL could be detected the same year, suggesting that several copies of the gene underlying the QTL are functional. The detection of some other homoeo-QTL was year-dependent. Therefore, changes in allelic expression could take place in response to environmental changes. We believe that, in strawberry as in other polyploid fruit species, the mechanisms unravelled in the present study may play a crucial role in the variations of fruit quality. Electronic supplementary material The online version of this article (doi:10.1007/s00122-011-1769-3) contains supplementary material, which is available to authorized users.

SummaryObjectives: Although graphical formats used to feedback clinical practice data may have an important impact, the most effective formats remain unknown. Using prevention of postoperative nausea and vomiting by anesthesiologists as an application, the objective of this study was to assess which graphical formats for feedback of clinical practice data are the most incentive to change practice.Methods: We conducted a multicenter cross-sectional study among anesthesiologists randomized in two groups between March andJune 2014. Each anesthesiologist assessed 15 graphical formats displaying an indicator of either prescription conformity or prescription effectiveness. Graphical formats varied by: type of graph (bar charts, linear sliders, or pictographs), presence or not of a target to reach, presence or not of a contrast between a hypothetical physician and his/her team, direction of the difference between the physician and his/her team, and restitution or not of the quality indicator evolution over the previous six months. The primary outcome was a numerical scale score expressing the anesthesiologists’ motivation to change his/her practice (ranging from 1 to 10 points). A linear mixed model was fitted to explain variation in motivation.Results: Sixty-six anesthesiologists assessed the conformity indicator and 67 assessed the effectiveness indicator. Factors associated with an increased motivation to change practice were: (i) presence of a clearly defined target to reach (conformity: β = 0.24 points, p = 0.0046; effectiveness: β = 1.11 points, p < 0.0001); (ii) contrast between the physician and his/her team (conformity: β = 0.38 points, p < 0.0001; effectiveness: β = 0.33 points, p = 0.0021); (iii) better results for the team than for the physician (conformity: β = 0.65 points, p < 0.0001; effectiveness β = 1.16 points, p < 0.0001). For the effectiveness indicator, anesthesiologists were more motivated to change practice with bar charts (β = 0.24 points, p = 0.0447) and pictographs (β = 0.45 points, p = 0.0001) than with linear sliders.Conclusions: Graphs associated with a defined target to reach should be preferred to deliver feedback, especially bar graphs or pictographs for indicators which are more complex to represent such as effectiveness indicators. Anesthesiologists are also more motivated to change practice when graphs report contrasted data between the physician and his/her team and a lower conformity or effectiveness for the physician than for his/her team.

Introduction: Mucus hypersecretion is one main pathophysiological mechanism of COPD. MUC5AC mucin is upregulated by cigarette smoke extract (CSE) in NCI-H292 cells cultures. Club Cell Secretory Protein (CCSP) is under-expressed in lung sections and bronchoalveolar lavage fluid of COPD patients. We already showed that CSE-induced IL-8 secretion was reduced after CCSP treatment in primary culture of human bronchial epithelial cells (HBEC) at air-liquid interface (ALI). We then hypothesized that whole cigarette smoke (WCS) would induce MUC5AC hyperproduction and that treatment with CCSP would reduce this hyperproduction. Materials and Methods: We stimulated both primary ALI cultures of HBEC from COPD, smoker and control patients, and NCI-H292 cells with WCS. We treated cells with ATP and/or CCSP. We assessed cell cytotoxicity via LDH activity. We determined MUC5AC level either by immunofluorescent staining of cultured cells, and by ELISA or Dot-Blot in cell lysates and supernatants. mRNA expression was assessed by RTqPCR. Results: In NCI-H292 cell line, viability was preserved in response to acute WCS exposure. We have shown an increase of IL-8 secretion but no MUC5AC protein using ELISA. In primary cultures of HBEC cell cytotoxicity tended to be altered thus not permitting us to show a significant modification of MUC5AC production. MUC5AC mRNA was overexpressed in COPD patients and seemed to be deacreased with CCSP treatment. Conclusions: No significant modification of MUC5AC production has been shown in primary ALI cultures of HBEC in response to acute WCS exposure and after CCSP treatment.

Objective The G8 is a screening test to identify frail elderly patients with cancer. Objectives were to design and evaluate the performance of alternative tests taking into account other predictive domains for frailty. Methods We conducted a literature review to identify predictive factors of frailty. Using a Delphi consensus, we collected 24 European experts' opinions to validate the most relevant items to improve the G8. Alternative tests were created and performance assessed on a development population (ONCODAGE cohort). The highest performing test was compared to the G8, and validated through both an internal and an external population validation (Aquitaine Geriatric Oncology cohort). Results The study population consisted of 1435 patients (ONCODAGE cohort) and 364 patients (Aquitaine Geriatric Oncology cohort). Twenty-three experts validated two items with a strong consensus (> 75%): modification of the threshold for the G8 polypharmacy item to six drugs per day and replacement of the G8 item on neuropsychological problems by four Instrumental Activities of Daily Living (IADL) items predictive of incident dementia, creating three modified G8 tests (addition of either item, or both). Only the G8 IADL-modified test had better performance than the G8 when tested on the ONCODAGE cohort: sensitivity = 77%, specificity = 67%. This test was validated on the internal (sensitivity = 78%, specificity = 71%) and external (sensitivity = 88%, specificity = 69%) validation populations. Conclusion Adding the four IADL items improves the performance of the G8. We have developed and validated a G8-modified test that is more specific than the G8 to detect frail elderly, while still sensitive and feasible in less than 10 min.

International audience; BACKGROUND Club cell secretory protein (CCSP) is a protective biomarker associated with annual decline in lung function. COPD progression results from an imbalance between injury and repair initially triggered by cigarette smoking. OBJECTIVE We investigated the effect of CCSP as a therapeutic strategy to restore the balance between injury and repair in COPD simultaneously, validating an ex vivo air-liquid interface (ALI) culture of human bronchial epithelial cells. METHODS Endobronchial biopsy specimens (EBBs) were obtained from 13 patients with COPD, eight smokers, and eight control subjects. Morphometric analysis of the initial EBBs was performed. ALI cultures derived from the same EBBs were exposed to cigarette smoke extract (CSE) with or without exogenous recombinant human CCSP (rhCCSP) supplementation. CCSP and IL-8 concentrations were assessed at steady state and after CSE exposure. RESULTS Morphometric analysis of the initial EBBs showed increased cell density but decreased immunostaining of CCSP+ cells in EBBs of patients with COPD (P = .03 vs control subjects). At steady state, lower CCSP (P = .04) and higher IL-8 levels (P < .0001) were found in COPD ALI epithelium. Exogenous rhCCSP supplementation dampened CSE-induced IL-8-release in patients with COPD and returned to levels similar to those of smokers and control subjects (P = .0001). A negative correlation was found between IL-8-release in ALI and CCSP+ cell density in initial biopsy specimens (P = .0073). CONCLUSIONS In vitro, rhCCSP exogenous supplementation can reverse CSE-induced IL-8 release in biopsy specimens from patients with COPD, indicating a potential use of this strategy in vivo.

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Introduction Les anticoagulants oraux (ACO) sont l’une des classes medicamenteuses les plus a risque de iatrogenie. La pertinence des prescriptions d’ACO constitue un enjeu majeur pour l’amelioration de la qualite et de la securite des soins. Devant le manque d’indicateurs de pertinence des prescriptions d’ACO, l’objectif de ce travail etait d’identifier des indicateurs de pertinence des prescriptions d’ACO en medecine adulte et d’evaluer leur utilite et leur caractere operationnel. Methodes Une revue de la litterature a ete realisee pour identifier des indicateurs de pertinence des prescriptions d’ACO a partir des recommandations issues des hautes autorites de sante ou societes savantes europeennes. Ces recommandations, publiees entre 2005 et 2015, portaient sur les prescriptions d’ACO en general ou pour des situations cliniques frequentes ciblees (fibrillation auriculaire, valvulopathies et protheses valvulaires, maladie thromboembolique veineuse, traumatismes ou chirurgie programmee). Une preselection des indicateurs a ete realisee par le comite scientifique, notamment sur des criteres de faisabilite. Un consensus de type Delphi modifie a ete mis en place pour evaluer l’utilite (capacite de l’indicateur a identifier des defauts de pertinence des prescriptions d’ACO et mettre en place des actions de correction) et le caractere operationnel en termes de frequence ou de gravite (capacite de l’indicateur a identifier des defauts de pertinence suffisamment frequents ou graves pour esperer une amelioration des pratiques) des indicateurs. Quarante experts europeens (France, Belgique et Suisse) specialises en cardiologie, medecine vasculaire, neurologie, geriatrie, medecine d’urgence, anesthesie-reanimation, pharmacologie ou hematologie ont ete selectionnes. Lors de chaque tour, les experts devaient coter l’utilite et le caractere operationnel sur une echelle binaire (oui ou non). Resultats A l’issue de la revue de litterature, 32 references ont ete analysees et ont permis d’identifier 84 indicateurs potentiels de pertinence des prescriptions d’ACO. Parmi ces indicateurs, 19 ont ete preselectionnes par le comite scientifique en vue de leur soumission pour jugement aupres d’experts. Parmi les experts sollicites, 22 experts (55 %) ont accepte de participer et ont repondu au premier tour du consensus Delphi. A l’issue du premier tour, 10 indicateurs ont ete retenus par obtention d’un consensus fort pour leur selection (≥ 80 % des experts les ayant juges a la fois utiles et operationnels en termes de frequence ou de gravite) ; aucun indicateur n’a ete rejete par obtention d’un consensus fort pour leur rejet (≥ 80 % des experts les ayant juges a la fois non utiles et non operationnels en termes de frequence ou de gravite) et 9 indicateurs n’ont pas obtenu de consensus. Ces 9 indicateurs ont ete resoumis pour jugement par les experts dans le cadre d’un second tour en cours. Conclusion Cette premiere etape de l’etude PACHA vise a repondre au manque actuel d’indicateurs de pertinence des prescriptions d’ACO, au plus pres des recommandations de pratiques et des indications cliniques validees. Les indicateurs juges les plus utiles et les plus operationnels a l’issue du consensus seront mis en œuvre a partir du systeme d’information hospitalier en vue de leur evaluation metrologique et de leur restitution aux prescripteurs dans le cadre d’une demarche structuree d’amelioration de la qualite et de la securite des soins.

IntroductionThe appropriateness of oral anticoagulant prescriptions is a major challenge to improve quality and safety of care. As indicators of the appropriateness of oral anticoagulant prescriptions are lacking, the aim of the study is to develop and validate a panel of such indicators, in hospitalised adults, from the hospital information system of two university hospitals in France.Methods and analysisThe study will be carried out in four steps: (1) a literature review to identify indicators of the appropriateness of oral anticoagulant prescriptions and their conditions of appropriateness; (2) a Delphi consensus method to assess the potential utility and operational implementation of the selected indicators; (3) techniques of medical data search to implement indicators from the hospital information system and; (4) a cross-sectional study to assess the ability of indicators to detect inappropriate oral anticoagulant prescriptions, performance of medical data search techniques for tracking or retrieving information and the ability of tools to be transferred into other institutions. The fourth step will include up to 80 patient hospital stays for each indicator, depending on the prevalence of inappropriate prescriptions estimated in interim analyses.Ethics and disseminationThis work addresses the current lack of quality indicators of the appropriateness of oral anticoagulant prescriptions. We aim to develop and validate such indicators for integrating them into hospital clinical practice, as part of a structured approach to improve quality and safety of care. As each hospital information system is different, we will propose tools transferable to other healthcare institutions to allow an automated construction of these indicators. The PACHA study protocol was approved by institutional review boards and ethics committees (CPP Sud-Ouest et Outre Mer III—DC 2016/119; CPP Ile-de-France II—CDW_2016_0014).Registration detailsClinical Trial.gov registration:NCT02898090.

Increasing evidence of a critical role played by the bronchial epithelium in airway homeostasis is opening new therapeutic avenues. Its unique situation at the interface with the environment suggests that the subtle regulation orchestrated by the epithelium between tolerance and specific immune response might be impaired in asthma. Airway mucus is acting as a physical and a biological fluid between the environment and the epithelium, synergistically moved by the cilia. In asthma, excessive mucus production is a hallmark of airway remodeling. Since many years we tried to therapeutically target mucus hypersecretion, but actually this option is still not achieved. The present review discusses the dynamic processes regulating airway mucus production. Airway inflammation is central in current asthma management. Understanding of how the airway epithelium influences the TH2 paradigm in response to deleterious agents is improving. The multiple receptors expressed by the airway epithelium are the transducers of the biological signals induced by various invasive agents to develop the most adapted response. Airway remodeling is observed in severe chronic airway diseases and may result from ongoing disturbance of signal transduction and epithelial renewal. Chronic airway diseases such as asthma will require assessment of these epithelial abnormalities to identify phenotypic characteristics associated with predicting a clinical benefit for epithelial-directed therapies.

International audience; Cold stress affects plant growth and development. In order to better understand the responses to cold (chilling or freezing tolerance), we used two contrasted pea lines. Following a chilling period, the Champagne line becomes tolerant to frost whereas the Terese line remains sensitive. Four suppression subtractive hybridisation libraries were obtained using mRNAs isolated from pea genotypes Champagne and Terese. Using quantitative polymerase chain reaction (qPCR) performed on 159 genes, 43 and 54 genes were identified as differentially expressed at the initial time point and during the time course study, respectively. Molecular markers were developed from the differentially expressed genes and were genotyped on a population of 164 RILs derived from a cross between Champagne and Terese. We identified 5 candidate genes colocalizing with 3 different frost damage quantitative trait loci (QTL) intervals and a protein quantity locus (PQL) rich region previously reported. This investigation revealed the role of constitutive differences between both genotypes in the cold responses, in particular with genes related to glycine degradation pathway that could confer to Champagne a better frost tolerance. We showed that freezing tolerance involves a decrease of expression of genes related to photosynthesis and the expression of a gene involved in the production of cysteine and methionine that could act as cryoprotectant molecules. Although it remains to be confirmed, this study could also reveal the involvement of the jasmonate pathway in the cold responses, since we observed that two genes related to this pathway were mapped in a frost damage QTL interval and in a PQL rich region interval, respectively.

International audience; Strawberry (Fragaria sp.) stands as an interesting model for studying flowering behaviour and its relationship with asexual plant reproduction in polycarpic perennial plants. Strawberry produces both inflorescences and stolons (also called runners), which are lateral stems growing at the soil surface and producing new clone plants. In this study, the flowering and runnering behaviour of two cultivated octoploid strawberry (Fragaria×ananassa Duch., 2n=8×=56) genotypes, a seasonal flowering genotype CF1116 and a perpetual flowering genotype Capitola, were studied along the growing season. The genetic bases of the perpetual flowering and runnering traits were investigated further using a pseudo full-sibling F1 population issued from a cross between these two genotypes. The results showed that a single major quantitative trait locus (QTL) named FaPFRU controlled both traits in the cultivated octoploid strawberry. This locus was not orthologous to the loci affecting perpetual flowering (SFL) and runnering (R) in Fragaria vesca, therefore suggesting different genetic control of perpetual flowering and runnering in the diploid and octoploid Fragaria spp. Furthermore, the FaPFRU QTL displayed opposite effects on flowering (positive effect) and on runnering (negative effect), indicating that both traits share common physiological control. These results suggest that this locus plays a major role in strawberry plant fitness by controlling the balance between sexual and asexual plant reproduction.

Background The majority of Non-Hodgkin’s lymphoma (NHL) patients are over 65 years. Management is challenging, especially for aggressive lymphoma, and appropriate assessment of efficacy and tolerance specific to this population is crucial. Objectives To assess the representation of older patients in randomised controlled trials (RCT) in NHL, examining whether trial eligibility criteria prevent participation, and whether appropriate primary endpoints such as toxicity, quality of life, or geriatric assessment scores are used. Methods We searched Medline for articles published in English or French between 1 January 2005 and 31 December 2011 reporting on phase II/III RCT evaluating therapeutic strategies for NHL. Articles were categorised as including or excluding (directly or indirectly) older adults, and features of RCT that included or excluded older patients are compared. Results We identified 87 relevant RCT: 9 (10.3%) focussed exclusively on patients >65 years, 22 (25.3%) directly excluded patients >65 years, 47 (54.0%) indirectly excluded older adults through selective inclusion criteria (ECOG status, liver or kidney function, and comorbidities), and 9 (10.3%) did not directly or indirectly exclude patients >65 years (although two excluded patients >70 years). Proportions of older patients included do not reflect incidence. Trials including older adults were published in journals with lower impact factors and few RCT used appropriate endpoints for older adults. Conclusions Older adults are poorly represented in NHL RCT both due to direct age-based exclusion and restrictive inclusion criteria. This situation needs rapid correction to better represent older patients and thus improve cancer management in this highly prevalent population.